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AB258387

Human CTSH (Cathepsin H) knockout HEK-293T cell lysate

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CTSH KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon3.

View Alternative Names

ACC 4, ACC 5, Aleurain, CATH_HUMAN, CPSB, CTSH, Cathepsin B3, Cathepsin BA, Cathepsin H heavy chain, Cathepsin H light chain, Cathepsin H mini chain, Minichain, N-benzoylarginine-beta-naphthylamide hydrolase, pro cathepsin H

2 Images
Sanger Sequencing - Human CTSH (Cathepsin H) knockout HEK-293T cell lysate (AB258387)
  • Sanger seq

Unknown

Sanger Sequencing - Human CTSH (Cathepsin H) knockout HEK-293T cell lysate (AB258387)

Homozygous : 11 bp deletion in exon3

Western blot - Human CTSH (Cathepsin H) knockout HEK-293T cell lysate (AB258387)
  • WB

Unknown

Western blot - Human CTSH (Cathepsin H) knockout HEK-293T cell lysate (AB258387)

Lanes 1-3 : Merged signal (red and green). Green - ab88960 observed at 27 kDa. Red - loading control ab181602 observed at 36 kDa.

ab88960 Anti-Cathepsin H antibody [AF14D7] was shown to specifically react with Cathepsin H in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266245 (knockout cell lysate ab258387) was used. Wild-type and Cathepsin H knockout samples were subjected to SDS-PAGE. ab88960 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Anti-Cathepsin H antibody [AF14D7] (ab88960) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

CTSH knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 3:

Western blot - Human CTSH (Cathepsin H) knockout HEK-293T cell lysate (ab258387) at 20 µg

Secondary

Lanes 1 - 3:

Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution

Lanes 1 - 3:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution

Observed band size: 37 kDa

false

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 11 bp deletion in exon3.

Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

What's included?

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Properties and storage information

Gene name
CTSH
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C

Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Cathepsin H also known as CPSH is a lysosomal cysteine protease that plays a role in intracellular protein degradation. It has a molecular mass of approximately 28 kilodaltons. This enzyme is expressed across various tissues with high expression levels in liver kidney and spleen. The enzyme primarily functions at acidic pH levels within lysosomes where it acts to break down peptides into amino acids by removing terminal residues.
Biological function summary

As a member of the cathepsin family Cathepsin H contributes to the regulation of protein turnover and homeostasis. It performs a dual role as both an aminopeptidase and endopeptidase. In addition to its function in protein degradation the enzyme involves itself in antigen processing during immune responses. Reactivity of Cathepsin H in immune complexes shows its importance in shaping immune tolerance and responses.

Pathways

Cathepsin H interacts with the lysosomal degradation pathway and plays a role in the MHC class II-mediated antigen presentation. This pathway is pivotal for the regulation of immune responses. Relation to other proteins like Cathepsin L and Cathepsin B highlights Cathepsin H's complementary function in proteolytic processing within lysosomes. Its role and expression play an integral part in balancing protein synthesis and degradation cycles.

Cathepsin H associates closely with cancer and neurodegenerative diseases. Its activity can influence tumor progression and metastasis due to its role in extracellular matrix remodeling and cell migration. In neurodegenerative disorders such as Alzheimer's disease its dysregulated activity may contribute to neuronal cell death. Cathepsin H’s connection to proteins like amyloid precursor protein (APP) in Alzheimer's highlights the complex interplay between protease activity and disease mechanisms.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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